Dynamical interplay between the human high-affinity copper transporter hCtr1 and its cognate metal ion

Gulshan Walke, Jana Aupič, Hadeel Kashoua, Pavel Janoš, Shelly Meron, Yulia Shenberger, Zena Qasem, Lada Gevorkyan-Airapetov, Alessandra Magistrato, Sharon Ruthstein

Research output: Contribution to journalArticlepeer-review

6 Scopus citations

Abstract

Abnormal cellular copper levels have been clearly implicated in genetic diseases, cancer, and neurodegeneration. Ctr1, a high-affinity copper transporter, is a homotrimeric integral membrane protein that provides the main route for cellular copper uptake. Together with a sophisticated copper transport system, Ctr1 regulates Cu(I) metabolism in eukaryotes. Despite its pivotal role in normal cell function, the molecular mechanism of copper uptake and transport via Ctr1 remains elusive. In this study, electron paramagnetic resonance (EPR), UV-visible spectroscopy, and all-atom simulations were employed to explore Cu(I) binding to full-length human Ctr1 (hCtr1), thereby elucidating how metal binding at multiple distinct sites affects the hCtr1 conformational dynamics. We demonstrate that each hCtr1 monomer binds up to five Cu(I) ions and that progressive Cu(I) binding triggers a marked structural rearrangement in the hCtr1 C-terminal region. The observed Cu(I)-induced conformational remodeling suggests that the C-terminal region may play a dual role, serving both as a channel gate and as a shuttle mediating the delivery of copper ions from the extracellular hCtr1 selectivity filter to intracellular metallochaperones. Our findings thus contribute to a more complete understanding of the mechanism of hCtr1-mediated Cu(I) uptake and provide a conceptual basis for developing mechanism-based therapeutics for treating pathological conditions linked to de-regulated copper metabolism.

Original languageEnglish
Pages (from-to)1194-1204
Number of pages11
JournalBiophysical Journal
Volume121
Issue number7
DOIs
StatePublished - 5 Apr 2022

Bibliographical note

Publisher Copyright:
© 2022 Biophysical Society

Funding

This research was funded by ERC -STG grant no. 754365 awarded to S.R. J.A. kindly acknowledges the Giovanni Fraviga Italian Association for Cancer Research (AIRC) fellowship. P.J. was supported by the HPC Europa Fellowship ( HPC17M882U ) under the project HPC-EUROPA3 (INFRAIA-2016-1-730897). A.M. thanks the Italian Association for Cancer Research (project AIRC IG 24514) and regional operational program ERDF 2014-2020 of the Friuli Venezia Giulia Region (project ARES - Against bRain cancEr: finding personalized therapies with in Silico and in vitro strategies) for financial support. This research was funded by ERC-STG grant no. 754365 awarded to S.R. J.A. kindly acknowledges the Giovanni Fraviga Italian Association for Cancer Research (AIRC) fellowship. P.J. was supported by the HPC Europa Fellowship (HPC17M882U) under the project HPC-EUROPA3 (INFRAIA-2016-1-730897). A.M. thanks the Italian Association for Cancer Research (project AIRC IG 24514) and regional operational program ERDF 2014-2020 of the Friuli Venezia Giulia Region (project ARES - Against bRain cancEr: finding personalized therapies with in Silico and in vitro strategies) for financial support.

FundersFunder number
ERC-STG
Giovanni Fraviga Italian Association for Cancer ResearchHPC17M882U, INFRAIA-2016-1-730897
Horizon 2020 Framework Programme754365
European Research Council
Associazione Italiana per la Ricerca sul CancroAIRC IG 24514
European Regional Development Fund
Regione Autonoma Friuli Venezia Giulia

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